Enabling high-sensitivity live single-cell mass spectrometry using an integrated electrical lysis and nano...

Area of Science:

• Single-cell analysis
• Metabolomics
• Mass Spectrometry

Background:

• Live single-cell metabolomics faces challenges with low sensitivity and limited metabolite detection due to small sample volumes.
• Current methods often omit cell lysis, leading to sample dilution and reduced analytical performance.
• Understanding cellular heterogeneity and responses to stimuli requires sensitive single-cell metabolic profiling.

Purpose of the Study:

• To develop an improved method for live single-cell metabolomics with enhanced sensitivity and metabolite coverage.
• To address the limitations of traditional methods by incorporating an efficient cell lysis step.
• To enable more accurate analysis of cellular responses and heterogeneity.

Main Methods:

• Development of an integrated single-cell electrical lysis and nano spray (SCEL-nS) platform.
• Coupling the SCEL-nS platform to an Orbitrap Mass Spectrometry (MS) system.
• Validation of lysis efficiency using live cell stain fluorescence and MS detection of tamoxifen-spiked cells.

Main Results:

• The SCEL-nS platform demonstrated efficient single-cell lysis, confirmed by reduced fluorescent intensity of live cell stains.
• MS analysis of single cells using SCEL-nS showed a statistically significant increase in measured peaks compared to non-lysis methods.
• The platform successfully detected metabolites from single cells, including those treated with tamoxifen.

Conclusions:

• The developed SCEL-nS platform significantly enhances sensitivity in live single-cell metabolomics.
• This method overcomes limitations of traditional approaches by incorporating efficient electrical lysis.
• SCEL-nS offers a promising advancement for detailed cellular analysis in various biological and medical fields.